Efficient derivation of stable pluripotent bovine embryonic stem cells
AssigneesThe Regents of the University of California, The Salk Institute for Biological Science
InventorsPablo Juan Ross, Yanina Bogliotti, Marcela Vilarino, Juan Carlos Izpisua Belmonte, Jun Wu
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Abstract
This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.
Claims26
A method of creating one or more ungulate animals comprising,
- obtaining one or more ungulate embryonic stem cells;
- producing one or more ungulate embryos from the one or more ungulate embryonic stem cells;
- and
- transferring the one or more ungulate embryos into one or more ungulate recipients to create one or more ungulate animals.
An isolated embryonic stem cell produced by a process comprising culturing an ungulate blastocyst cell or a pluripotent cell isolated from an ungulate embryo, in a cell culture media,
- wherein the cell culture media comprises:
- (i) an organic matrix, in replacement of inactivated feeder cells;
- (ii) Activin A;
- (iii) an effective amount of Fibroblast Growth Factor 2 (FGF2) or an equivalent thereof;
- and (iv) an effective amount of one or more inhibitors of Wnt signaling.
A population of isolated embryonic stem cell produced by a process comprising culturing and expanding an ungulate blastocyst cell or a pluripotent cell isolated from an ungulate embryo, in a cell culture media,
- wherein the cell culture media comprises:
- (i) an organic matrix, in replacement of inactivated feeder cells;
- (ii) Activin A;
- (iii) an effective amount of Fibroblast Growth Factor 2 (FGF2) or an equivalent thereof;
- and (iv) an effective amount of one or more inhibitors of Wnt signaling.
An ungulate germ cell produced by a process comprising
- generating an ungulate embryonic stem cell (ESC), and
- in vitro differentiation of the ungulate ESC to germ cells by addition/subtraction of growth factors and/or co-culture with somatic cells.
Dependencies are inferred from the claim text.